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Published on: July 10, 2020
Identification and characterization of MAVS from black carp Mylopharyngodon piceus
Wei Zhou1, Jujun Zhou1, Ying Lv1
1Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, College of Life Science, Hunan Normal University, Changsha, 410081, China.
Abstract:
MAVS (mitochondria antiviral signaling protein) plays an important role in the host cellular innate immune response against microbial pathogens. In this study, MAVS has been cloned and characterized from black carp (Mylopharyngodon piceus). The full-length cDNA of black carp MAVS (bcMAVS) consists of 2352 nucleotides and the predicted bcMAVS protein contains 579 amino acids. Structural analysis showed that bcMAVS is composed of functional domains including an N-terminal CARD, a central proline-rich domain, a putative TRAF2-binding motif and a C-terminal TM domain, which is similar to mammalian MAVS. bcMAVS is constitutively transcribed in all the selected tissues including gill, kidney, heart, intestine, liver, muscle, skin and spleen; bcMAVS mRNA level in intestine, liver, muscle increased but decreased in spleen right after GCRV or SVCV infection. Multiple bands of bcMAVS were detected in western blot when it was expressed in tissue culture, which is similar to mammalian MAVS. Immunofluorescence assay determined that bcMAVS is a mitochondria protein and luciferase reporter assay demonstrated that bcMAVS could induce zebrafish IFN and EPC IFN expression in tissue culture. Data generated in this manuscript has built a solid foundation for further elucidating the function of bcMAVS in the innate immune system of black carp.
Insights
Researchers characterized the black carp MAVS (mitochondria antiviral signaling protein), revealing its crucial role in innate immunity. This study provides foundational data for understanding MAVS function in fish antiviral defense.
Area of Science:
- Immunology
- Molecular Biology
- Fish Science
Background:
- Mitochondria antiviral signaling protein (MAVS) is critical for innate immune responses against pathogens.
- Understanding MAVS in teleost fish is essential for aquaculture health and disease management.
Purpose of the Study:
- To clone and characterize the MAVS gene from black carp (Mylopharyngodon piceus).
- To investigate the expression patterns and subcellular localization of black carp MAVS (bcMAVS).
- To assess the functional role of bcMAVS in inducing antiviral gene expression.
Main Methods:
- Full-length cDNA cloning and sequence analysis of bcMAVS.
- Quantitative analysis of bcMAVS mRNA expression in various tissues and after viral challenge (GCRV, SVCV).
- Western blot, immunofluorescence assay, and luciferase reporter assay to determine protein expression, localization, and function.
Main Results:
- The bcMAVS gene was successfully cloned, encoding a 579-amino acid protein with conserved functional domains similar to mammalian MAVS.
- bcMAVS is constitutively expressed in multiple tissues and its expression is modulated by viral infections.
- bcMAVS localizes to mitochondria and can induce interferon (IFN) expression in zebrafish and EPC cells.
Conclusions:
- Black carp MAVS shares structural and functional similarities with its mammalian counterparts.
- bcMAVS plays a significant role in the innate immune response of black carp against viral pathogens.
- This study lays the groundwork for further research into the specific mechanisms of bcMAVS-mediated antiviral immunity in fish.
